The microbiota associated with the human vagina exists in a mutualisfic relafionship with the human host and is believed to play an important role in women's reproducfive health. The vaginal microbial communities constitute the first line of defense against infection by invasive non-indigenous organisms that cause disease, such as the sexually transmitted Chlamydia trachomatis. Despite their importance, surprisingly litfie is known about the composition and dynamics of vaginal microbial communifies in health and disease. Traditional culfivafion-based methods have provided a valuable but incomplete picture of the human vaginal microbiota. In this study, we will combined massively-parallel sequencing technology with the culture-independent analysis of the 16S rRNA gene sequence to survey the vaginal microbiota species composifion and abundance in young adults with C. trachomatis infecfion and C. frac/70/r7af/s-positive women with pelvic inflammatory disease (PID). In addifion, we will establish the dynamics of the community a subgroup of women sampled longitudinally over one year or more after treatment. In each of these women, we will use community transcriptomics to identify the suite of genes expressed by the vaginal microbial community. This combined data will afford a unique view of the vaginal microbiota dynamics during and after Chlamydial infecfion (i.e., a detailed picture of the metabolic pathways triggered in response to the infecfions (direcfiy or indirectly)], and will further our model of Chlamydial infecfion and re-ihfecfion. Because of limitations in using humans as research subjects, guinea pigs are used as animal model for Chlamydial infections. Similarlly, we will characterize the vaginal microbiota in healthy and C. cawae-infected female guinea pigs over time. Understand and characterizing the importance of the vaginal microbiota will contribute greafiy to the development of new approaches based on rafionale and scienfifically sound principles to manipulate the vaginal microbiota in parallel to treatments. The genome of more than 200 C.trachomatis or C. caviae isolated from these biological samples will be sequenced using 454 pyrosequencing. These sequences will represent an unparalleled resource that will be shared with the research community. The sequence data will be analyzed in correlafion with the vaginal microbiota and the phenotypes characterized under the two projects of this consortium.